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. 1997 Aug 11;138(3):517–529. doi: 10.1083/jcb.138.3.517

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Analysis of API maturation in vam3^tsf cells. Wild-type (SEY6210) cells and TDY1 (vam3Δ) cells carrying a vam3^tsf plasmid (pVAM3-6.414) were incubated at 38°C for 5 min, labeled with [³⁵S]cysteine/methionine for 10 min, and then chased for the indicated times. Equivalent volumes of labeled culture were harvested at the indicated time points of chase. API was recovered from lysates by immunoprecipitation, subjected to SDS-PAGE, and analyzed by autoradiography. The cytoplasmic precursor (pr) and mature vacuolar (m) forms of API are indicated.